CN107446897A - A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof - Google Patents

A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof Download PDF

Info

Publication number
CN107446897A
CN107446897A CN201710675790.7A CN201710675790A CN107446897A CN 107446897 A CN107446897 A CN 107446897A CN 201710675790 A CN201710675790 A CN 201710675790A CN 107446897 A CN107446897 A CN 107446897A
Authority
CN
China
Prior art keywords
crude enzyme
enzyme liquid
fish liver
freeze
liver crude
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201710675790.7A
Other languages
Chinese (zh)
Inventor
王海燕
王碧燕
韩大雄
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Xiamen University
Third Institute of Oceanography SOA
Original Assignee
Xiamen University
Third Institute of Oceanography SOA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Xiamen University, Third Institute of Oceanography SOA filed Critical Xiamen University
Priority to CN201710675790.7A priority Critical patent/CN107446897A/en
Publication of CN107446897A publication Critical patent/CN107446897A/en
Pending legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/25Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving enzymes not classifiable in groups C12Q1/26 - C12Q1/66
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90Enzymes; Proenzymes
    • G01N2333/902Oxidoreductases (1.)
    • G01N2333/90283Oxidoreductases (1.) acting on superoxide radicals as acceptor (1.15)
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90Enzymes; Proenzymes
    • G01N2333/91Transferases (2.)
    • G01N2333/91194Transferases (2.) transferring sulfur containing groups (2.8)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention discloses fish liver crude enzyme liquid freeze-dried powder and its preparation method and application.The fish liver crude enzyme liquid freeze-dried powder is the dry powder product of fish liver crude enzyme liquid.The present invention discloses the preparation method of the fish liver crude enzyme liquid freeze-dried powder, and the fish liver crude enzyme liquid of brand-new is prepared into fish liver crude enzyme liquid freeze-dried powder with long preservation period by freeze drier.The fish liver crude enzyme liquid freeze-dried powder of the present invention can apply to the measure of the Yi Fen azolactones ethoxyresorufin O-deethylase of 7 ethyoxyl 3, glutathione S-transferase, the detection of superoxide dismutase isoreactivity and reduced glutathione equal size.

Description

A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof
Technical field
The invention belongs to biochemical field, and in particular to a kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof and should With.
Background technology
Microsome be cell by homogenate it is broken when, the folliculus that oneself is reclosed after the membrane structure rupture of endomembrane system Bubble (mainly endoplasmic reticulum), diameter about 100nm or so of these vesicles, is heterogeneous aggregate, referred to as micro- Plastochondria.Microsome mainly contains cytochrome P 450 Enzyme (CYP) and antioxidant system albumen.Fish liver crude enzyme liquid is fish liver The supernatant obtained after the broken centrifugation of homogenate, is mainly made up of microsome, containing a variety of biotransferases and albumen, such as 7- ethoxies Base -3- Yi Fen azolactones ethoxyresorufin O-deethylases (7-ethoxyresorufin-o-deethylase, EROD), glutathione sulphur-transfer Enzyme (glutathione S-transferase, GST), superoxide dismutase (superoxide dismutase, SOD) and Glutathione (glutathione, GSH) etc..Research finds Cytochrome P450 (such as:EROD etc.) and antioxidant system enzyme system (such as:GST, SOD etc.) and polyphenoils is (such as:GSH etc.) to pollutant (such as heavy metal, brominated flame-retardant, polycyclic in environment Aromatic hydrocarbons, Polychlorinated biphenyls etc.) there is certain response relation.Existing many scholars by detect the EROD in fish liver crude enzyme liquid, The change of the active and GSH of the enzymes such as GST, SOD etc. content carrys out indicative for environments situation, but fish liver crude enzyme liquid is liquid condition, Be inconvenient to store with quantitative, and with the extension of holding time, easy inactivation, and enzyme source is used as using fish liver crude enzyme liquid freeze-dried powder There is not been reported for the research of progress enzymatic activity and GAP-associated protein GAP assay.
The content of the invention
It is an object of the invention in place of overcome the deficiencies in the prior art, there is provided fish liver crude enzyme liquid freeze-dried powder and its system Preparation Method and application.
One of the technical solution adopted for the present invention to solve the technical problems is:
Fish liver crude enzyme liquid freeze-dried powder is obtained dry powder product after the freeze-drying of fish liver crude enzyme liquid.
Wherein, the fish liver crude enzyme liquid freeze-dried powder is the dry powder system obtained after brand-new fish liver crude enzyme liquid is freeze-dried Product.
The two of the technical solution adopted for the present invention to solve the technical problems are:
The preparation method of above-mentioned fish liver crude enzyme liquid freeze-dried powder, including:
1) cut open after fish takes out liver and (avoid staving fish courage), washed with 0.10~0.30% KCl solution of precooling, filter paper Sop up bloodstain.Weigh, by 1:5 (W/V) ratio adds the PBS (pH 7.0~8.0) of precooling, is homogenized under ice bath.Homogenate is 0 At~8 DEG C, 8000~12000g centrifuges 10~30min, takes supernatant to dispense.
2) by the fish liver crude enzyme liquid being prepared in -80~-40 DEG C of refrigerators or liquid nitrogen cryopreservation 24~72 hours, to thick enzyme Liquid fully charge.
3) the fish liver crude enzyme liquid freeze drier of freezing is dried 24~72 under -80~-40 DEG C and vacuum condition Hour to fish liver crude enzyme liquid is changed into dry dry powder-shaped, in -20~-4 DEG C of preservations.
The three of the technical solution adopted for the present invention to solve the technical problems are:
Above-mentioned fish liver crude enzyme liquid freeze-dried powder is in 7- ethyoxyl -3- Yi Fen azolactones ethoxyresorufin O-deethylase, glutathione sulphur-transfer Application in the measure of enzyme, the detection of superoxide dismutase isoreactivity and glutathione equal size.
Compared with background technology, it has the following advantages that the technical program:
1. the invention provides fish liver crude enzyme liquid freeze-dried powder and preparation method thereof.
2. of the invention dried brand-new fish liver crude enzyme liquid by freeze drier under -80~-40 DEG C and vacuum condition It is changed into dry dry powder-shaped to fish liver crude enzyme liquid within 24~72 hours, for traditional fish liver crude enzyme liquid, its dry powder Form is more convenient storage than the form of liquid, quantified, and stability is preferable, avoids the fish liver crude enzyme liquid of traditional freezen protective Influence of the frozen-thaw process to enzymatic activity during use.
3. the fish liver crude enzyme liquid freeze-dried powder of the present invention can be used for 7- ethyoxyl -3- Yi Fen azolactones ethoxyresorufin O-deethylase, paddy The measure of the sweet peptide sulfurtransferase of Guang, the detection of superoxide dismutase isoreactivity and glutathione equal size.
Brief description of the drawings
The invention will be further described with reference to the accompanying drawings and examples.
Fig. 1 is the fish liver crude enzyme liquid freeze-dried powder sample drawing obtained in embodiment.
Fig. 2 is EROD enzyme of the fish liver crude enzyme liquid with fish liver crude enzyme liquid freeze-dried powder of the different batches obtained in experimental example Expression activitiy, wherein abscissa is fish liver enzyme source batch, and ordinate is the activity of EROD enzymes.
Embodiment
Present disclosure is illustrated below by embodiment.
Embodiment:Prepare fish liver crude enzyme liquid freeze-dried powder
Cut open after fish (Tilapia mossambica) takes out liver and (avoid staving fish courage), washed with 0.15% KCl solution of precooling, filter paper Sop up bloodstain.Weigh, be 13.8g, shred, by 1:5 (W/V) ratio adds PBS (pH 7.4) 69.0mL of precooling, under ice bath It is homogenized 3min.Homogenate 10000g centrifugation 20min, takes supernatant to dispense at 4 DEG C.The fish liver crude enzyme liquid that will be prepared It is placed in liquid nitrogen cryopreservation 24 hours.The fish liver crude enzyme liquid freeze drier of freezing is dried 36 under -80 DEG C and vacuum condition Hour, dried powder, as fish liver crude enzyme liquid freeze-dried powder are taken out, is placed in -20 DEG C of preservations.
The fish liver crude enzyme liquid freeze-dried powder sample drawing being prepared is as shown in Figure 1.
Above-described embodiment can realize the technique effect of following experimental examples:
Experimental example 1:The EROD enzyme assays of fish liver crude enzyme liquid freeze-dried powder
(1) preparation of samples:10mg fish liver crude enzyme liquid freeze-dried powders are taken, 50mg/mL is dissolved into 200 μ L pH 7.5 PBS Crude enzyme liquid.
(2) active testing:At room temperature, crude enzyme liquid 20 μ L, 0.01mol/L PBS (pH 7.5) 160 are added in 96 orifice plates μ L, 0.5mmol/L the μ L of 7- ethyoxyls -3- Yi Fen azolactones (ERF) solution 5, it is eventually adding 0.5mmol/L NADPH solution 40 μ L, is mixed immediately using 532nm as excitation wavelength, using 580nm as launch wavelength, determine fluorescence intensity.Determined once per 30s Fluorescent value, METHOD FOR CONTINUOUS DETERMINATION 25min.
7- ethyoxyl -3- Yi Fen azolactone ethoxyresorufin O-deethylase the activity measured is:0.59pmol/min/mg pro
Experimental example 2:The EROD activity ratios of different batches fish liver crude enzyme liquid and fish liver crude enzyme liquid freeze-dried powder compared with
(1) the EROD enzyme assays in different batches fish liver crude enzyme liquid:Operation is with reference to the step of experimental example 1 (2).
(2) the EROD enzyme assays in different batches fish liver crude enzyme liquid freeze-dried powder:Operation is with reference to the step of experimental example 1 (1)、(2)。
The experimental result of the EROD enzyme assays of different batches fish liver crude enzyme liquid and fish liver crude enzyme liquid freeze-dried powder is such as Shown in Fig. 2.
Above-mentioned experimental result is shown:The EROD enzymatic activitys of fish liver crude enzyme liquid freeze-dried powder are unlike thick enzyme made from each batch Liquid it is active low, substantially freeze-dried powder activity for crude enzyme liquid activity 77-120%, illustrate the freeze-dried powder storage new as crude enzyme liquid The form of depositing has feasibility.
Experimental example 3:The GST enzyme assays of fish liver crude enzyme liquid freeze-dried powder
10mg fish liver crude enzyme liquid freeze-dried powders are taken, 50mg/mL crude enzyme liquid is dissolved into 200 μ L pH 6.5 PBS.Room Under temperature, crude enzyme liquid 10 μ L, 0.01mol/L PBS (pH 6.5) 170 μ L, 30mmol/L GSH solution 10 are added in 96 orifice plates μ L, the 30mmol/L μ L of CDNB solution 10 are eventually adding, mixed.Non- enzymatic reaction is not added with enzyme liquid, immediately per 30s under 340nm Determine a light absorption value, METHOD FOR CONTINUOUS DETERMINATION 5min.
The GST enzymatic activitys measured are:15U/mL
Experimental example 4:The SOD enzyme activity measure of fish liver crude enzyme liquid freeze-dried powder
10mg fish liver crude enzyme liquid freeze-dried powders are taken, 50mg/mL crude enzyme liquid is dissolved into 200 μ L pH 8.2 PBS.Room Under temperature, the 0.01mol/L μ L of Tris-HCl solution (pH 8.2) 100 are added in 96 orifice plates, the μ L of ultra-pure water 90, are eventually adding The 4.5mmol/L μ L of pyrogallol solution 10, mix and determine a light absorption value per 30s at 325 nm immediately, determine 5min.Meter Mouse thymus cells speed is calculated, the μ L of supernatant 10 is added by above step, autoxidation speed is suppressed about 50%.
The SOD enzyme activity measured is:110U/mL
Experimental example 5:The GSH assays of fish liver crude enzyme liquid freeze-dried powder
10mg fish liver crude enzyme liquid freeze-dried powders are taken, 50mg/mL crude enzyme liquid is dissolved into 200 μ L pH 8.0 PBS.Room Under temperature, 10 μ L crude enzyme liquids, 180 μ L 0.01mol/L phosphate buffer solution (pH 8.0) and 10 μ L neighbour's benzene are added in 96 orifice plates The ethanol solution (1mg/mL) of dicarbaldehyde, fully mix, 20min. is placed at room temperature using 340nm as excitation wavelength, under 430nm Determine fluorescence intensity.
The GSH measured content is:0.8mmol/L
It is described above, preferred embodiments only of the present invention, therefore the scope that the present invention is implemented can not be limited according to this, i.e., according to this The equivalent changes and modifications that patent of invention scope and description are made, it should all still fall within the range of the present invention covers.

Claims (4)

1. fish liver crude enzyme liquid freeze-dried powder, it is characterised in that:The liver crude enzyme liquid freeze-dried powder is that the freezing of fish liver crude enzyme liquid is dry Dry powder product after dry.
2. fish liver crude enzyme liquid freeze-dried powder according to claim 1, it is characterised in that:The fish liver crude enzyme liquid freeze-dried powder The dry powder product obtained after being freeze-dried for brand-new fish liver crude enzyme liquid.
3. the preparation method of fish liver crude enzyme liquid freeze-dried powder according to claim 1, it is characterised in that:
1) after cuing open fish taking-up liver, washed with 0.10~0.30% KCl solution of precooling, filter paper sops up bloodstain;Weigh, by 1: 5 (W/V) ratio adds the PBS (pH 7.0~8.0) of precooling, is homogenized under ice bath;Homogenate at 0~8 DEG C, 8000~ 12000g centrifuges 10~30min, takes supernatant to dispense;
2) it is complete to crude enzyme liquid by the fish liver crude enzyme liquid being prepared in -80~-40 DEG C of refrigerators or liquid nitrogen cryopreservation 24~72 hours Freeze entirely;
3) the fish liver crude enzyme liquid freeze drier of freezing is dried 24~72 hours under -80~-40 DEG C and vacuum condition It is changed into dry dry powder-shaped to fish liver crude enzyme liquid, in -20~-4 DEG C of preservations.
4. fish liver crude enzyme liquid enzyme dry powder according to claim 1 is taking off ethyl including 7- ethyoxyl -3- Yi Fen azolactones The detection of enzymatic activity including enzyme, glutathione S-transferase, superoxide dismutase and reduced glutathione content Purposes in measure.
CN201710675790.7A 2017-08-09 2017-08-09 A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof Pending CN107446897A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710675790.7A CN107446897A (en) 2017-08-09 2017-08-09 A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710675790.7A CN107446897A (en) 2017-08-09 2017-08-09 A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof

Publications (1)

Publication Number Publication Date
CN107446897A true CN107446897A (en) 2017-12-08

Family

ID=60491715

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710675790.7A Pending CN107446897A (en) 2017-08-09 2017-08-09 A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof

Country Status (1)

Country Link
CN (1) CN107446897A (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002062969A9 (en) * 2001-02-06 2002-10-17 Massachusetts Inst Technology Cellular reprogramming in peptide hydrogel and uses thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002062969A9 (en) * 2001-02-06 2002-10-17 Massachusetts Inst Technology Cellular reprogramming in peptide hydrogel and uses thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
曾青兰 等: "《生物制药工艺》", 29 February 2012, 华中科技大学出版社 *
朱智 等: "EROD、GST测试法在评价污染物生态效应的研究", 《中国科技论文在线》 *

Similar Documents

Publication Publication Date Title
Testino Jr et al. High-throughput inhibition screening of major human cytochrome P450 enzymes using an in vitro cocktail and liquid chromatography–tandem mass spectrometry
Li et al. Comparison of the pro-oxidative and proinflammatory effects of organic diesel exhaust particle chemicals in bronchial epithelial cells and macrophages
Chen et al. Age-related increase in mitochondrial superoxide generation in the testosterone-producing cells of Brown Norway rat testes: relationship to reduced steroidogenic function?
Olinga et al. Comparison of five incubation systems for rat liver slices using functional and viability parameters
Hildebrandt et al. Mechanism of coumarin action: sensitivity of vitamin K metabolizing enzymes of normal and warfarin-resistant rat liver
Piper et al. Preadaptation to efficient respiratory maintenance is essential both for maximal longevity and the retention of replicative potential in chronologically ageing yeast
Clarke et al. Characterization of the inhibition of P4501A2 by furafylline
Chou et al. Thermotolerance of isolated mitochondria associated with heat shock proteins
Brito et al. Resveratrol affords protection against peroxynitrite-mediated endothelial cell death: A role for intracellular glutathione
Tibaldi et al. Src‐Tyrosine kinases are major agents in mitochondrial tyrosine phosphorylation
González et al. Characterization of aromatase activity in the sea bass: effects of temperature and different catalytic properties of brain and ovarian homogenates and microsomes
Vitali et al. Properties and catalytic activities of MICAL1, the flavoenzyme involved in cytoskeleton dynamics, and modulation by its CH, LIM and C-terminal domains
Ranjan et al. Direct effects of neuropeptide nesfatin-1 on testicular spermatogenesis and steroidogenesis of the adult mice
Wang et al. Glutathione regulates the transfer of iron-sulfur cluster from monothiol and dithiol glutaredoxins to apo ferredoxin
Pham et al. Cadmium-induced apoptosis in rat hepatocytes does not necessarily involve caspase-dependent pathways
Salganicoff et al. Energy metabolism of blood platelets: I. Isolation and properties of platelet mitochondria
Stapelfeldt et al. Menadione-mediated WST1 reduction assay for the determination of metabolic activity of cultured neural cells
CN107446897A (en) A kind of fish liver crude enzyme liquid freeze-dried powder and preparation method thereof
Hsieh et al. Involvement of reactive oxygen species in PTTH-stimulated ecdysteroidogenesis in prothoracic glands of the silkworm, Bombyx mori
Joseph-Horne et al. Characterization of a split respiratory pathway in the wheat “take-all” fungus, Gaeumannomyces graminis var. tritici
Xie et al. Effect of glucose levels on carbon flow rate, antioxidant status, and enzyme activity of yeast during fermentation
Modriansky et al. Human hepatocyte-A model for toxicological studies. Functional and biochemical characterization
Sims et al. Purification and characterization of the isoprene monooxygenase from Rhodococcus sp. strain AD45
Chai et al. Mechanistic insight into allosteric activation of human pyruvate carboxylase by acetyl-CoA
Seglen et al. Tryptophan oxygenase activation and ascorbate oxidation in whole homogenates from rat liver

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20171208